Intermediates useful in camptothecin synthesis

ABSTRACT

The present invention provides a process for the asymmetric synthesis of camptothecin analogues as well as novel chemical intermediates of Formula I, II, and III. In general, the present process comprises conversion of a cis dioxolanone, having the same desired absolute configuration as the desired camptothecin analogue, to a compound of Formula I, II, or III, which compound is then converted to the desired camptothecin analogue. ##STR1##

This is a continuation application of application Ser. No. 08/363,509,filed Dec. 23, 1994 now abandoned, which is a divisional application ofSer. No. 08/075,063, filed Jun. 10, 1993, now U.S. Pat. No. 5,405,963.

FIELD OF THE INVENTION

The present invention relates to chemical processes and novel chemicalintermediates which are useful for preparing pharmaceutically importantcamptothecin analogues such as (S)-10-(dimethylamino)methyl!-4-ethyl-4,9-dihydroxy-1H-pyrano3',4':6,7!indolizino 1,2-b!quinoline-3,14(4H, 12H)dionemonohydrochloride, commonly known as topotecan; or(S)-4-ethyl-4,9-dihydroxy- 1H-pyrano 3'4':6,7!indolizino1,2-b!quinoline-3,14(4H,12H)dione, commonly known as10-methoxycamptothecin, in high enantiomeric excess.

BACKGROUND OF THE INVENTION

Camptothecin analogues, such as topotecan, have been shown to be usefulas both antineoplastic and antiviral therapeutic agents. A process forthe total synthesis of racemic topotecan, a camptothecin analogue, isdescribed in copending U.S. Ser. No. 07/941,496, now abandoned. A recentexample of a different total synthesis was published by Comins, et al.,J. Am. Chem. Soc., 114, 10971, 1992. As is often the case, optimaltherapeutic activity is provided by only one configuration of themolecule. It is therefore desirable to produce this material in a formwhich is highly enriched in only one absolute configuration of thechiral center.

A. I. Meyers et al., J. Org. Chem., 38, 1973, 1974 describes thesynthesis of racemic camptothecin analogues by the coupling of acarboxylic acid with a pyrrolo 3,4-b!quinoline. However, that processlacks the desirable ability to introduce chirality into the targetcamptothecin analogue.

SUMMARY OF THE INVENTION

An object of this invention is to provide novel chemical intermediateswhich are useful in the asymmetric synthesis of camptothecin analogues.

Another object of the present invention is to provide processes forpreparing chemical intermediates useful in the asymmetric synthesis ofcamptothecin analogues.

In one aspect, this invention provides compounds according to Formulae(I), (II) or (III): ##STR2## wherein: X is selected from a groupconsisting essentially of a cyano group; a carboxylic acid; an N-arylcarboxylic acid derivative of Formula (A); ##STR3## and a good leavinggroup, preferably a halide not including chloride, yet more preferably abromide or iodide, most preferably bromide; or a sulfonate, morepreferably a p-fluorobenzenesulfonate, trifluoromethanesulfonate orfluorosulphonate, most preferably trifluoromethanesulfonate;

Y is selected from a group consisting essentially of a cyano group; acarboxylic acid; an N-aryl carboxylic acid derivative of Formula (A);##STR4## and a good leaving group, preferably a halide, more preferablya chloride, bromide or iodide, most preferably bromide; or a sulfonate,more preferably a p-fluorobenzenesulfonate, trifluoromethanesulfonate orfluorosulphonate, most preferably trifluoromethanesulfonate;

R is propargyl (i.e., --CH₂ C.tbd.CH) or substituted propargyl, as inFormula (B); ##STR5## R₁ is hydrogen, or R₁ together with R₂ is --OCH₂O--; R₂ is hydrogen; O(C₁ -C₆) alkyl, preferably methoxy; OH; or R_(l)together with R₂ is --OCH₂ O--;

R₃ is hydrogen; OH; O(C₁ -C₆) alkyl; NO₂ ;NH₂ ; or a protected nitrogengroup which can be convened to NH₂, preferably trifluoroacetamido,acetamido, methoxycarbonylamino or carboxenzyloxyamino;

R₄ is hydrogen; SiMe₃ ; or alkyl, preferably methyl or ethyl;

R₅ is hydrogen or COOR₆ ;

R₆ is a carboxylic acid ester group, preferably methyl, ethyl, n-propyl,isopropyl, n-butyl, s-butyl, t-butyl, allyl or benzyl esters; and

R₇ and R₈ are both hydrogen or R₇ together with R₈ is CH(t-butyl).

In another aspect, this invention provides novel methods for theasymmetric total synthesis of camptothecin analogues.

DETAILED DESCRIPTION OF THE INVENTION

Known methods for synthesizing camptothecin analogues primarily resultin racemic products. However, the antiviral or antineoplastic activityof camptothecin analogues has been observed to vary with enantiomericpurity. That is, one enantiomer of a given analogue exhibits superiorantiviral or antineoplastic activity when compared with that of theother enantiomer. For example the unnatural enantiomer of camptothecin(R) exhibits cytotoxicity without having the specific anticanceractivity associated with the naturally-occurring enantiomer (S). Thecompounds of the present invention, represented by the Formulae (I),(II), and (III): ##STR6## wherein: X is selected from a group consistingessentially of a cyano group; a carboxylic acid; an N-aryl carboxylicacid derivative of Formula (A); ##STR7## and a good leaving group,preferably a halide not including chloride, more preferably a bromide oriodide, most preferably bromide; or a sulfonate, more preferably ap-fluorobenzenesulfonate, trifluoromethanesulfonate or fluorosulphonate,most preferably trifluoromethanesulfonate;

Y is selected from a group consisting essentially of a cyano group; acarboxylic acid; an N-aryl carboxylic acid derivative of Formula (A);##STR8## and a good leaving group, preferably a halide, more preferablya chloride, bromide or iodide, most preferably bromide; or a sulfonate,more preferably a p-fluorobenzenesulfonate, trifluoromethanesulfonate orfluorosulphonate, most preferably trifluoromethanesulfonate;

R is propargyl (i.e., --CH₂ C.tbd.CH) or substituted propargyl, as inFormula (B); ##STR9## R₁ is hydrogen, or R₁ together with R₂ is --OCH₂O--; R₂ is hydrogen; O(C₁ -C₆) alkyl, preferably methoxy; OH; or R₁together with R₂ is --OCH₂ O;

R₃ is hydrogen; OH; O(C₁ -C₆) alkyl; NO₂ ;NH₂ ; or a protected nitrogengroup which can be converted to NH₂, preferably, trifluoroacetamido,acetamido, methoxycarbonylamino or carboxenzyloxyamino;

R₄ is hydrogen; SiMe₃ ; or alkyl, preferably, methyl or ethyl;

R₅ is hydrogen or COOR₆ ;

R₆ is a carboxylic acid ester group, preferably methyl, ethyl, n-propyl,isopropyl, n-butyl, s-butyl, t-butyl, allyl or benzyl esters; and

R₇ and R₈ are both hydrogen or R₇ together with R₈ is CH(t-butyl),

are useful as chemical intermediates in the production of camptothecinanalogues by the asymmetric syntheses of the present invention.

The following terms have the meanings defined below here and throughoutthis application.

The term "good leaving group" means any chemical moiety which one ofordinary skill in the art would understand to be sufficientlyelectrophilic so as to readily undergo nucleophilic substitution in thepresence of a nucleophile, for example chloride, bromide, iodide,p-fluorobenzenesulfonate, trifluoromethylsulfonate or fluorosulphonate.

The term "protected nitrogen group which can be converted to NH₂ "includes any chemical moiety which one of ordinary skill in the artwould understand to be the protected equivalent of --NH₂ and capable ofreadily being deprotected by known methods to --NH₂, preferablytrifluoroacetamido, acetamido, methoxycarbonylamino orcarboxenzyloxyamino.

The term "alkyl" includes common C₁ -C₆ saturated hydrocarbon chainssuch as methyl, ethyl, n-propyl, isopropyl, n-butyl, s-butyl andt-butyl.

The term "camptothecin" includes camptothecin and any derivative thereofthe structure of which is based on the 1H-pyrano 3',4':6,7!indolizino-1,2-b!quinolin-3,14(4H, 12H)-dione ring system.

The terms "1H-pyrano 3'4':6,7:!quinoline", "indolizino1,2-b!quinolin-9(11H)-one" and "1H-pyrano 3',4':6,7!indolizino-1,2-b!quinolin-3,14(4H, 12H)-dione" refer generally to compounds basedon these ring systems.

The term "camptothecin analogue" includes camptothecins as defined aboveand also includes derivatives of camptothecin wherein the E ring hasbeen replaced with another functionality.

The term "carboxylic acid" refers to --COOH.

The term "cyano group" refers to C.tbd.N.

The term "carboxylic acid ester groups" refers to methyl, ethyl,n-propyl, isopropyl, n-butyl, s-butyl, t-butyl, allyl and benzyl esters.

The term "strong alkylating agent" includes agents capable of producingeither an O-alkylimidate or imidate ester, imidoyl halide, or imidederivative in the process of the present invention.

The following section discloses how to make the intermediate compoundsof the present invention and also discloses the processes of the presentinvention. The first process of the present invention is illustrated inSchemes 1-4. In general this process comprises:

a) forming a chiral cis dioxolanone having a carboxylic acid function,the cis dioxolanone having the absolute configuration desired in thecamptothecin analogue;

b) forming a pyrrolo 3,4-b!quinoline having a free amine function;

c) reacting the cis dioxolanone with the pyrrolo 3,4-b!quinoline to forma compound of Formula (III) ##STR10## wherein: R₁ is hydrogen or R₁together with R₂ is --OCH₂ O--; preferably hydrogen;

R₂ is hydrogen; O(C₁ -C₆)alkyl, preferably methoxy; OH; or R₁ togetherwith R₂ is --OCH₂ O--;

R₃ is hydrogen; OH; O(C₁ -C₆)alkyl; NO₂ ;NH₂ ; or a protected nitrogenwhich can be converted to NH₂, preferably hydrogen;

R₄ is hydrogen; SiMe₃ ; or alkyl, preferably hydrogen or alkyl, the mostpreferred alkyl being ethyl;

R₅ is hydrogen or COOR₆ ;

R₆ is a carboxylic acid ester group, preferably methyl, ethyl, n-propyl,isopropyl, n-butyl, s-butyl, t-butyl, allyl or benzyl esters, mostpreferably methyl ester; and

R₇ and R₈ together form CH(t-butyl); and

d) converting the compound of Formula (III) into a camptothecinanalogue.

Schemes 6 and 7 illustrate the second process of the present invention.In general this process comprises:

a) forming a chiral cis dioxolanone having the absolute configurationdesired in the camptothecin analogue;

b) converting the cis dioxolanone to a compound of Formula (I) ##STR11##wherein X is selected from a group consisting of a good leaving group,preferably a halide not including chloride, more preferably a bromide oriodide; or a sulfonate, more preferably, p-fluorobenzenesulfonate,trifluoromethanesulfonate or fluorosulphonate; a cyano group; acarboxylic acid; and an N-aryl carboxylic amide derivative of Formula(A); ##STR12## c) converting said compound of Formula (I) into acompound of Formula (II) ##STR13## wherein: R is propargyl orsubstituted propargyl of Formula (B); ##STR14## Y is selected from agroup consisting of a good leaving group, a cyano group, a carboxylicacid, and an N-aryl carboxylic amide derivative of Formula (A); and##STR15## d) converting the compound of Formula (II) into a camptothecinanalogue. Both processes rely on the use of a chiral 1,3 dioxolan-4-oneto introduce the desired absolute configuration into the desiredcamptothecin analogue product.

Chiral 1,3-dioxalan-4-one is prepared from commercially availableR-2-aminobutyric acid. Diazotisation of the amino acid with sodiumnitrite and sulfuric acid is accompanied by the displacement of thediazo group with complete retention of configuration to giveR-2-hydroxybutyric acid. Similar reactions have been described by K.Mori, et al. in Tetrahedron, 35, 1601, 1979. The chiral hydroxyl-bearingcarbon eventually becomes the chiral center (C-20) of the camptothecinmolecule. This hydroxyacid is converted to the corresponding acetal, (1)in Scheme I, with a very high stereoselectivity (greater than 20:1) forformation of the cis dioxolanone. This specific reaction was disclosedby K. Krohn, et al. in Annalen Der Chemie., 949, 1988, although we havemodified these authors' reaction conditions to achieve an improvedstereoselectivity over their reported results (improved from about10-14:1). Recrystallization of the dioxolanone at low temperature (-70°C.) from hexane:ether gives the essentially pure (>99%) cis isomer. Thisstep allows control of the absolute stereochemistry of the chiral centerof camptothecin, since all subsequent chemical operations are carriedout with retention of the configuration at this carbon. This approach tocontrol of absolute stereochemistry is useful in the preparation ofcamptothecin and its analogues in essentially complete enantiomericpurity by either of the two processes of the present invention.##STR16##

Step a) of the first process described as follows. Scheme 1 illustratesa process for making the dioxolanone starting material. As shown inScheme 2 Knoevenagel condensation of the commercially availablemono-methyl acetal of glyoxaldehyde with methylbenzylmalonate givesmethylene malonate (2). Subsequent Michael addition of an enolate,preferably the lithium enolate, of (1) gives compound (3), in which thestereochemistry of the dioxolanone is as shown. Refer to, D. Seebach, etal. in "Modern Synthetic Methods, 1986" R. Scheffold ed. pages 125-259for a discussion of the control of the stereoselectivity observed in theproduct. This reaction establishes the absolute configuration of thechiral center, eventually leading to the desired (S)-camptothecins,since all subsequent operations are carried out without affecting thestereochemical integrity of this center. Hydrogenolysis of the benzylester produces the corresponding acid (4). ##STR17##

In step b) of the first process of the present invention, thesubstituted pyrrolo 3,4-b!quinoline (6) is prepared as shown in Scheme3. Para-anisidine is acylated on nitrogen using bromoacetyl bromide (oroptionally, chloroacetyl chloride). The remaining, alkyl bromide (orchloride) group is subsequently displaced in an SN₂ reaction usingpropargylamine, and the secondary amine nitrogen is protected by formingthe corresponding carbamate derivative with methyl chloroformate toyield (5). Compound (5) is reacted in a polar solvent, preferablymethylene chloride, 1,2-dichloroethane, 1,2-dimethoxyethane,tetrahydrofuran, N,N-dimethylformamide, acetonitrile, acetone orN-methylpyrrolidinone, most preferably acetonitrile; at a startingtemperature of about -30° C., followed by warming to a temperature ofabout 20°-85° C., preferably at 40°-85° C., most preferably at atemperature of 40°-50° C.; in the presence of a strong alkylating agentor such agent as is capable of producing either an O-alkylimidate orimidate ester, imidoyl halide, or such imide derivative defined hereinand throughout this application as "imidate/imidoyl halide derivative";preferably trifluoromethanesulfonic anhydride, dimethylsulfate,alkyloxonium tetrafluoroborates, aluminum chloride,O-benzyltrichloroacetimidate, triphenylphosphine/carbon tetrachloride,or triphenylphosphine/carbon tetrabromide, most preferablytrimethyloxonium tetrafluoroborate. The resulting imidate/imidoyl halidederivative then undergoes 4+2!cycloaddition and subsequent eliminationof methanol (in the case in which an O-methyl imidate ester is initiallyformed) or hydrogen halide (in the case in which an imidoyl halide isinitially formed) or a sulfonic acid (in the case in which an imidateester is initially formed) to yield a substituted pyrrolo3,4-b!quinoline (6). This transformation has also been described in U.S.Ser. No. 07/941,496.

In step c) of the first process of the present invention, the carboxylicacid functionality of the cis dioxolanone is reacted with the pyrrolo3,4-b!quinoline at the amine function to form an amide having a dimethylacetal function. This dimethyl acetal function of the amide is thenhydrolyzed to form an aldehyde. Intramolecular aldol condensation ofthis aldehyde, followed by aromatization, yields a compound of Formula(III). ##STR18##

In particular, amine (7) is prepared from (6) by hydrolysis of thecarbamate function with acetic acid saturated with hydrobromic acid asshown in Scheme 4. This intermediate is coupled with the carboxylic acid(4) prepared as in Scheme 2 by utilizing1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (abbreviated WSCD for"water-soluble carbodiimide" in the Scheme) and hydroxybenzotriazole(abbreviated HOBT) to give the resulting amide (8). The dimethyl acetalfunction is hydrolyzed with boron trifluoride etherate to give theresulting aldehyde (9). Aldol condensation using trifluoroacetic acid topromote the elimination of water, with subsequent aromatisation, using2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) as an oxidizing agent:gives the aromatic D-ring skeleton of the camptothecins shown, with(10a) as the major product. This product is an intermediate compound ofthe present invention of Formula (III).

In step d) of the first process of the present invention, thisintermediate of Formula (III), wherein R₅ is methyl ester, undergoesreduction at the methyl ester to an alcohol function, followed byintramolecular lactone formation to form a camptothecin analogue. Forexample, subsequent reduction of the methyl ester of (10a) to thecorresponding alcohol and closure of the E ring lactone by basichydrolysis of the dioxolanone ring and subsequent acidification gives anasymmetric total synthesis of (S)-10-methoxycamptothecin (11). ##STR19##

Alternatively, in step c) of the first process, reacting a cisdioxolanone with a pyrrolo 3,4-b!quinoline also results in a product ofFormula (III), wherein R₅ is hydrogen. In general, this compound isconverted to a camptothecin analogue by a process comprising:

1) forming a Mannich reagent known as1,1,3,3-tetramethyl-diaminomethane;

2) reacting the Mannich reagent with the compound of Formula (III),wherein R₅ is hydrogen, in a reaction mixture to form aN,N-dimethylaminomethylated intermediate;

3) treating the reaction mixture with an aqueous base; and

4) acidifying the reaction mixture to precipitate a camptothecinanalogue.

This process is exemplified in Scheme 4 in which compound (10b) isisolated as a byproduct of the reaction/isolation sequence used toprepare (10a). This product arises from (10a) by decarbomethoxylation,apparently during chromatographic isolation. Compound (10b) is alsoeasily converted to (S)-10-methoxycamptothecin using a novel processwhich yields a significant improvement upon existing knowledge. Thehydroxymethylation of compound (A) shown in Scheme 5 was reported byDanishefsky et al., J. Am. Chem. Soc., 94, 5576, 1972 to proceed upontreatment of the substrate with paraformaldehyde in dioxane containingconcentrated sulfuric acid (100 ° C., 16 hours) to yield the precursorof racemic camptothecin which lacks only the required hydroxyl group atC-20. However, this is a difficult reaction to carry out, typicallygiving the desired product in only a low (approximately 25%) yield. Theprocess of the present invention provides a greatly improved conversionof compound (10b) to (S)-10-methoxycamptothecin. By utilizing areactive, pre-formed Mannich reagent(1,1,3,3-tetramethyl-diaminomethane), compound (10b) can be very readilyN,N-dimethylaminomethylated (rather than hydroxymethylated). Thisintermediate is not isolated (refer to Scheme 4) but rather worked up bysequential treatment with aqueous base to displace dimethylamine andconcurrently hydrolyze the acetal of the dioxolanone ring. This isfollowed by acidification to close the lactone ring and precipitate thedesired product, (S)-10-methoxycamptothecin in very good (65%) overallyield. ##STR20##

A second process of the present invention for the synthesis ofcamptothecins possessing the natural enantiomeric configuration can alsobe executed utilizing chiral dioxolanone (1). This process, which isoutlined in Schemes 6 and 7, is substantially different from theabove-described process of the present invention. The chiral cisdioxolanone having a diester function described in step a) of the secondprocess can be formed by Michael addition of an enolate of dioxolanoneto diethyl glutaconate. Specifically, the dioxolanone enolate deprivedfrom (1) is added in a Michael fashion (1,4-addition) to diethylglutaconate to give diester (12).

In step b) of the second process, the cis dioxolanone is converted to acompound of Formula (I), wherein X is preferably a good leaving group,more preferably chloride, bromide or iodide, most preferably bromide, byhydrolysis of the diester function of the dioxolanone, followed by ringclosure to form a corresponding glutarimide and treatment of theglutarimide with an excess of a thionyl halide, preferably thionylbromide, in dimethylformamide, followed by workup in aqueous base. Thediester is then in this fashion hydrolyzed and ring-closed to give thecorresponding glutarimide (13) by treatment with ammonia in acetic acid.The glutarimide is then treated with excess thionyl chloride or thionylbromide in the presence of N,N-dimethylformamide. Workup with aqueousbase, followed by neutralization results in the formation of bicyclicintermediate (14a). While not wishing to be bound to any particularmechanism of action, it is believed that this step proceeds by thefollowing mechanism. Thionyl halide converts N,N-dimethylformamide intothe corresponding N,N-dimethyliminium halide (Vilsmeier reagent) whichserves to formylate the glutarimide ring at a position adjacent to oneof the carbonyl groups. The other carbonyl group of the glutarimide isconverted in the same process to the corresponding vinyl halide by theaction of excess thionyl halide. Workup with a basic aqueous solutionthen generates the isolated product. During the course of workup aprototropic shift occurs, causing rearrangement of the exocyclicaldehyde function to give the corresponding hydroxymethyl pyridone. Thehydroxymethyl group closes onto the carbonyl group of the chiraldioxolanone, ultimately resulting in loss of pivalaldehyde and formationof what will become the E ring lactone of camptothecin with an absoluteconfiguration identical to that possessed by the natural product.

Alternatively, compound (13), may be treated with a powerfulsulfonylating agent such as trifluoromethanesulfonic anhydride (triflicanhydride) in the presence of N,N-dimethylformamide, resulting in aproduct (14b) which is a vinyl sulfonate analogue of vinyl halides(14a). ##STR21##

A preferred process of the present invention, which provides for thetotal synthesis of (S)-10-methoxycamptothecin, is further described inthe Example below.

In the following synthetic Example, temperature is in degrees Centigrade(°C.). Unless otherwise indicated, all of the starting materials wereobtained from commercial sources and used as received. It is believedthat one skilled in the an can, using the preceding description, utilizethe present invention to its fullest extent. This Example is givenmerely to illustrate the present invention, and should not be construedas limiting the scope thereof in any way. Reference is made to theclaims for what is reserved to the inventor hereunder.

EXAMPLE

The following Example describes a preferred process of the presentinvention for the total synthesis of (S)-10-methoxycamptothecin, whichprocess is outlined in Schemes 1-4.

EXAMPLE 1 ##STR22##

(R)-2-Hydroxybutyric acid is prepared from (R)-2-aminobutyric acidfollowing the procedure of K. Mori et al. described in Tetrahedron, 35,1601, 1979. The reaction product was obtained in approximately 65% yieldwhen carried out on a scale utilizing 50 g of the starting aminobutyricacid. The (R)-2-aminobutyric acid was purchased from the AldrichChemical Corp. with a purity of 99%. The chiral purity of the desired(R)-2-hydroxybutyric acid was previously established by Mori as >97%.##STR23##

A solution of (R)-2-hydroxybutyric acid (7.2 g, 60 mmol) and2,2-dimethylpropionaldehyde (11.91 g, 138 mmol, 2.3 eq) in 200 ml ofpentane was treated with 250 mg (1.2 mmol, 2 mol % of p-toluenesulfonicacid and two drops (approximately 50 mg) of concentrated (12M) aqueoussulfuric acid solution. The solution was heated at reflux for 18 hourswith removal of water. The solution was cooled to ambient temperature,washed with 2×100 ml of water and dried over anhydrous magnesiumsulfate. The solution was filtered and concentrated under reducedpressure to 11.0 g of a yellow oil. This material was distilled at apressure of 50 mm Hg to give a main fraction (9.77 g, 82.0% yield) withboiling point of 93°-95° C. The product was examined by ¹ H NMRspectroscopy and determined to be an approximately 28:1 mixture of cis-and trans-dioxolanone ring isomers by comparison of the relativeintegrations of the methine singlets representing the --CH(tert-butyl)proton for the respective isomers.

The mixture of cis and trans dioxolanones (8.0 g) was dissolved indiethyl ether (about 25 ml) with subsequent cooling in an ice/salt bathto approximately -20° C. The solution was gradually diluted withstirring by the addition of hexane which was precooled to -20° C.Addition was continued until the solution became: slightly turbid(approximately 100 ml added). At this point a small amount of additionaldiethyl ether was added to give a clear solution, and stirring wascontinued with gradual cooling to -78° C. using dry ice/acetone. Aprecipitate of fine, white crystals was seen to form upon cooling. Theproduct was collected by rapid decanting of the cold solution, washingwith about 25 ml of -78° C. hexane, and filtration at -78° C. using achilled, jacketed Buchner funnel. The collected product was quicklyremoved for storage and was observed to melt at approximately -5° C. Thepresence of the minor, trans isomer of the dioxolanone ring was nolonger detectable using ¹ H NMR spectroscopy.

The chirality of the product was determined by ¹ H NMR spectroscopyusing a chiral shift reagent Eu(thd)₃,(tris(2,2,6,6-tetramethyl-3,5-heptanedionato)europium). A sample ofracemic material was prepared from racemic 2-hydroxybutyric acid andrecrystallized at low temperature as described above for the chiralproduct in order to prepare a sample for comparison. ##STR24##

A mixture of 4.16 g (20 mmol,) of benzyl methyl malonate, 5.20 g (50mmol, 2.5 eq) of glyoxal-1,1-dimethyl acetal, 1.7 g (20 mmol, 1.0 eq) ofpiperidine and 200 ml of toluene was refluxed with azeotropic removal ofwater using a Dean Stark trap. After 45 minutes the theoretical amountof water had been collected and the mixture was cooled to ambienttemperature. The yellow solution was evaporated under reduced pressureand chromatographed on silica gel using methylene chloride as eluent.The fractions containing product were evaporated to give 4.82 g of thedesired compound as a colorless oil (82.0% yield). ##STR25##

A 2.5M solution of n-butyllithium in hexane (1.6 ml, 4.0 mmol) was addedto a stirred, -78° C. solution of dry diisopropylamine (426 mg, 4.22mmol) in 40 ml of fleshly distilled tetrahydrofuran under an atmosphereof dry nitrogen. The mixture was warmed to 0° C. and stirred for about30 minutes. The pale yellow solution was then re-cooled to approximately-78° C. with continued stirring.

The dioxolanone (1) prepared as described in Step 1 (765 mg, 4.4 mmol)was dissolved in 5 ml of fleshly distilled tetrahydrofuran and cooled to-78° C. in a separate reaction vessel under an atmosphere of drynitrogen. This solution was then carefully transferred to the fleshlyprepared solution of lithium diisopropylamide via a double-ended needleover about 10 minutes. The temperature of the lithium diisopropylamidesolution was not allowed to rise above about -75° C. during the courseof the addition. The reaction solution was stirred for 45 minutes afterthe addition was complete. During this time a solution of compound (2)was prepared in 5 ml of dry tetrahydrofuran and cooled to -78° C., againunder an atmosphere of dry nitrogen. The solution of (2) was addeddropwise over about 10 minutes to the stirred enolate solution of (1).After stirring for approximately 3 hours with gradual warming to -20° C.the mixture was quenched by pouring into 30 ml of a half-saturatedsolution of aqueous ammonium chloride and extracted with 2×50 mlportions of diethyl ether. The combined organic layers were dried overanhydrous sodium sulfate, filtered and evaporated to give 1.70 g of aclear, colorless oil.

The product was chromatographed on 50 g of silica gel using 20% ethylacetate:hexane as eluent. After evaporation of solvent from theproduct-containing fractions, a total of 1.08 g (58.1%) of desiredproduct was obtained as a heavy, colorless oil. ##STR26##

A solution of 1.8 g (3.8 mmol) of the benzyl ester in 50 ml of absolutemethanol was treated with 250 mg of 5% palladium-on-carbon catalyst andreduced under an atmosphere of hydrogen at 50 psi for 1.75 hours.Hydrogen uptake was essentially complete after about 1 hour. The mixturewas vented to nitrogen and purged under an inert atmosphere. Thesuspension was filtered through a short pad of filter cel with suctionto remove the suspended catalyst, and evaporated under vacuum. Thecolorless oil was dried under a high vacuum (<0.001 mm Hg) to give 1.44g of a colorless oil (99% yield). ##STR27##

A solution of freshly distilled bromoacetyl bromide (20.22 g, 100 mmol)in 200 ml of methylene chloride was cooled with stirring to -23° C. Tothis was added a slight excess (11.1 g, 110 mmol) of triethylamine. Asolution of p-anisidine (12.3 g, 100 mmol) in 100 ml of methylenechloride was slowly added to the mixture with rapid stirring. A heavy,white precipitate formed upon the addition of the p-anisidine solution.The addition was exothermic and was carried out so that the internaltemperature of the solution did not exceed -10° C. Stirring wascontinued for 1.5 hours after addition was complete. The mixture wasdiluted with water (200 ml) and subsequently washed with an additional200 ml of water and 200 ml of 3% aqueous, hydrochloric acid solution.The methylene chloride layer was dried over anhydrous sodium sulfate,evaporated to a yellow-green solid, and triturated with 150 ml oftert-butyl methyl ether to give a white, crystalline powder afterfiltration. After drying under reduced pressure, 22.5 g (92.3%) of thedesired product was obtained. ##STR28##

A solution of 11.0 g (200 mmol) of propargylamine in 150 ml of methylenechloride was stirred at ambient temperature. To this was added 12.2 g(50 mmol) of the solid bromide in a single portion. Stirring wascontinued for a total of about 18 hours, at which time the reaction wascomplete as judged by analytical thin layer chromatography. The solutionwas washed with water (100 and 50 ml portions) and dried over anhydroussodium sulfate. The solution was filtered and evaporated under reducedpressure to yield an orange oil which crystallized under high vacuum toyield 10.77 g of crude product as orange crystals. The product waschromatographed by filtration through 70 g of silica gel using 5%methanol:methylene chloride as eluent. After evaporation of solvent fromthe product-containing fractions the desired compound was isolated byevaporation of solvent to yield 10.0 g of a pale-orange, crystallinesolid (91.7%). ##STR29##

A stirred solution of the secondary propargylamine (2.18 g, 10 mmol) andpyridine (790 mg, 10 mmol) in methylene chloride (25 ml) was treated bythe dropwise addition of 950 mg (10 mmol, 1.0 eq) of methylchloroformate at ambient temperature. The addition was mildlyexothermic. The mixture was stirred for a total of about 2 hours, atwhich time analytical thin layer chromatography indicated that thereaction was complete. The reaction was worked up by the addition ofwater (25 ml) and additional methylene chloride (25 ml). The organiclayer was washed with additional water (2×25ml) and dried over anhydroussodium sulfate. After filtration and evaporation, the resulting oil waschromatographed on silica gel (60 g) using 3% methanol:methylenechloride as eluent. The suitable fractions were combined and evaporatedto give 1.75 g (63.4% yield) of the desired product as a white solid.##STR30##

A 100 mg (0.36 mmol) sample of the N-propargyl urethane in solution with3.0 ml of dry acetonitrile was stirred under an atmosphere of nitrogenat -30° C. To this was added dropwise a solution of 160 mg (1.08 mmol,3.0 eq) of trimethyloxonium fluoroborate in 3.0 ml of dry acetonitrile.The addition was carried out over about 5-10 minutes so as to maintainthe internal temperature of the solution at -30° C. After stirring forone hour at -30° C. and one hour at ambient temperature, the mixture washeated to 50° C. and maintained at this temperature for 24 hours. Atthis point the solution was cooled to ambient temperature and thesolvent was removed under reduced pressure. The residual oil waspartitioned between 25 ml each of methylene chloride and water, and theaqueous phase was extracted with 3 additional 25 ml portions ofmethylene chloride. The combined organic extracts were washed with 50 mlof water, dried over anhydrous sodium sulfate, and evaporated to a brownsolid. The solid was recrystallized from methanol to give 155 mg ofproduct (60% yield) which was collected in two crops. ##STR31##

A solution of 1.00 g (3.9 mmol) of the carbamate in 20 ml of acetic acidsaturated with hydrobromic acid was refluxed for three hours. Duringthis time an insoluble, white precipitate formed in the solution. Themixture was allowed to stand overnight at ambient temperature tocomplete the precipitation. The product was isolated by filtration togive 1.45 g (100%) of product after drying under vacuum at 40° C. for 24hours to a constant weight. ##STR32##

A solution of 863 mg (2.4 mmol) of tricyclic amine dihydrobromide in 30ml of dry tetrahydrofuran was stirred with 485 mg (4.8 mmol, 2.0 eq) oftriethylamine and 507 mg (2.6 mmol, 1.1 eq) of1-(3-dimethylaminopropyl)-3-ethylcarbodiimide at ambient temperatureunder a nitrogen atmosphere. The progress of the reaction was monitoredby HPLC using a C₁₈ μBondapak column (50% acetonitrile:water containing0.1M sodium perchlorate adjusted to a pH of 2.5 by addition ofperchloric acid; detection by UV at a wavelength of 218 nm, flow rate2.0 ml/min; r.t. of starting amine 1.33 min., r.t. of product 9.88min.). After 24 hours the starting material had been essentiallyconsumed and the relative peak-area-ratio response for the major productwas 93.5%. The reaction was worked up by evaporating the solvent underreduced pressure to a brown oil. The residual material was partitionedbetween 50 ml each of methylene chloride and water. The aqueous layerwas extracted with an additional 50 ml of methylene chloride and thecombined organic extracts were dried over anhydrous sodium sulfate,filtered and evaporated to yield 1.5 g of a brown, glassy foam. Thisresidue was chromatographed through silica gel (60 g) using 10%methanol:methylene chloride as eluent. The suitable fractions werecombined and evaporated to give 1.0 g of product (74.7% yield) as ayellow oil. ##STR33##

A 500 mg sample of the starting acetal (0.89 mmol) was stirred in asolution of 50 ml of methylene chloride at -78° C. This was treated with0.8 ml (923 mg, 6.5 mmol, 7.3 eq) of boron trifluoride etherate added ina single portion. The solution was stirred for 20 minutes with continuedcooling, then stirred with the cooling bath removed for an additional 20minutes. The solution was diluted with 25 ml of water, then stirredvigorously for an additional 30 minutes. The pH of the solution was thenadjusted to approximately 6.0 by the addition of solid sodiumbicarbonate. Stirring was stopped, and the layers of the reaction wereseparated. The organic layer was dried over anhydrous sodium sulfate,filtered and evaporated to yield 500 mg (450 mg theoretical yield| of alight-colored foam. NMR spectroscopy confirmed that the dimethyl acetalof the starting material had been hydrolyzed to produce thecorresponding aldehyde.

The crude aldehyde was dissolved in 35 ml of toluene containing twodrops (about 40 mg) of trifluoroacetic acid. The mixture was refluxedfor 18 hours, washed with 25 ml of a saturated solution of aqueoussodium bicarbonate and evaporated to give 500 mg of a dark oil. About460 mg of this crude product was then carried on by dissolution in 45 mlof toluene. The toluene solution was stirred under an atmosphere ofnitrogen at ambient temperature, and 460 mg (1.98 mmol, 2.2 theoreticalmol eq) of 2,3-dichloro-5,6-dicyanobenzoquinone (DDQ) was added. Afterstirring for 18 hours the reaction was evaporated to a dark oil, andchromatographed on 100 g of silica gel using 10% methanol:methylenechloride as eluent. Approximately 300 mg of a dark oil was obtained. Aportion (approximately one-half) of this crude product was furtherpurified by an additional chromatography on silica gel (30 g). Afterelution with ethyl acetate to remove non-polar impurities, the productwas eluted with 5% methanol:ethyl acetate to yield 35 mg of product. Anadditional chromatography of this sample was carried out to yield 18 mgof pure product as determined by analytical TLC/HPLC. In addition to theisolation of (10a) from this chromatography, a second compound (9 mg,subsequently identified as compound (10b)) was also isolated as abyproduct. ##STR34##

An 11 mg (0.022 mmol) sample of chromatographed ester was dissolved in 1ml of methylene chloride and stirred under an atmosphere of nitrogen atambient temperature. The solution was treated with dropwise addition of0.1 ml (0.11 mmol, 5.0 eq) of diisobutylaluminum hydride added as a 1Msolution in hexane. After stirring for 1 hour, the mixture waspartitioned between water (2 ml) and methylene chloride (8 ml). Theorganic layer was dried over anhydrous sodium sulfate, filtered andevaporated to a heavy oil. Proton NMR spectroscopy indicated that thereduction of the methyl ester to the corresponding alcohol was notcomplete. A sample of appoximately 8 mg of the residue was thenre-dissolved in 2 ml of methylene chloride and cooled to -10° C. Thiswas treated with an additional 0.1 ml (0.11 mmol) of diisobutylaluminumhydride and stirred with slow warming to ambient temperature over about3 hours. The mixture was evaporated to a heavy oil and dissolved in 3 mlof absolute methanol. A solution of 2 N aqueous sodium hydroxide (1 ml)was added and the mixture was stirred for about 14 hours at ambienttemperature. The solution was neutralized by the addition of 500 mg ofglacial acetic acid mid evaporated to dryness under reduced pressure.The residue was dissolved in 1 ml of 1: 1 methylene chloride:methanoland purified by preparative thin layer chromatography using a silica gelmatrix. After elution twice with 10% methanol:methylene chloridecontaining 1% glacial acetic acid the product was isolated as 4.6 mg ofa pale-yellow solid (54.5% yield).

The identity and purity of the product was confirmed by comparison ofits proton nuclear mass resonance spectrum (¹ H NMR) mass spectrumincluding high-resolution exact mass determination (MS) andchromatographic behavior utilizing high pressure liquid chromatography(HPLC) versus an authentic sample of the desired material from naturalplant sources. The chirality of the product was determined by HPLCchromatography utilizing a commercially available column packed with achiral support medium.

¹ H NMR (DMSO; resonances quoted in ppm downfield fromtetramethylsilane): δ5 0.88 (t, 3H; J=7.2 Hz), 1.88 (m, 2H), 3.32 (br s,due to the presence of HOD), 3.93 (s, 3H), 5.25 (s, 2H), 5.50 (s, 2H),6.51 (s, 1H), 7.23 (s, 1H), 7.49 (d, 1H; J=7.5 Hz), 8.06 (d, 1H; J=7.5Hz), 8.54 (s, 1H)

Mass Spectrum (DCI/NH₃ ionization): m/z 379 (M+H)⁺ ; 335 (M-CO₂ +H)⁺

Exact Mass: calculated: 378.12158 found: 378.12159

HPLC conditions

Column: Two concatenated Techocel OA-3 100 columns (total dimensions 50cm ×4.0 mm)

Mobile Phase: 95:5 n-Butyl chloride:Methanol

Temperature: Ambient

Detector: UV at a wavelength of 370 nm

Flow Rate: 1.0 ml/minute

Injection Volume: 10 μl

Sample Run: 75 minutes

Sample Prep'n.: Sample is first dissolved in a minimum ofN,N-dimethylformamide, followed by diluting to an approximateconcentration of 0.1 mg/ml in the mobile phase.

Results of HPLC Analysis

Ret. Time: 41.61 minutes for (R) 10-methoxycamptothecin

45.11 minutes for (S) 10-methoxycamptothecin

Response: 99.1% total peak-area-ratio response for the major compound,(S) 10-methoxycamptothecin

0.55% total peak-area-ratio response for the opposite enantiomer, (R)10-methoxycamptothecin ##STR35##

A 16 mg sample (0.037 mmol) of solid, crystalline starting material wasstirred with 3 ml of glacial acetic acid at 20° C. to give a clear,pale-yellow solution. To this was added 15 mg (0.15 mmol, 4.0 eq) of1,1,3,3,-tetramethyldiaminomethane in a single portion. After stirringfor 1.5 hours at ambient temperature, the reaction mixture was heated to50° C. After stirring for 12 hours, analytical TLC confirmed that nostarting material remained in solution. The solution was thenconcentrated under vacuum at ambient temperature to an oil. The oil wasthen stirred for 2 hours at ambient temperature with 3 ml of a 2 Naqueous, sodium hydroxide solution. The aqueous solution was carefullyacidified to a pH of approximately 3.5-4.0 by the dropwise addition of5% aqueous, hydrochloric acid solution. A solid was seen to precipitatefrom the aqueous solution. The aqueous suspension was diluted to avolume of 15 ml and extracted with 3×35 ml portions of 20%methanol:methylene chloride. The organic layers were then combined andevaporated to dryness under reduced pressure. The residual solid wastriturated with 5 ml of hot methanol and the product was isolated byfiltration. The identity of the product (9.0 mg, 65% yield) wasconfirmed in the same manner as for the immediately preceding reaction.

What we claim:
 1. A compound of Formula (II): ##STR36## wherein R ispropargyl or substituted propargyl of Formula (B); ##STR37## wherein R₄is hydrogen, SiMe₃ or alkyl;Y is selected from a group consistingessentially of, bromo, iodo, a sulfonate, a cyano group, a carboxylicacid, and an N-aryl carboxylic amide derivative of Formula (A) ##STR38##wherein R₁ is hydrogen, or R₁ together with R₂ is --OCH₂ O--; R₂ ishydrogen, O(C₁ -C₆) or OH; and R₃ is hydrogen, OH, O(C₁ -C₆)alkyl, NO₂,NH₂, trifluoroacetamido, acetamido or methoxycarbonylamino.
 2. Acompound according to claim 1 wherein R is propargyl.
 3. A compoundaccording to claim 1 where Y is bromo, iodo, or a sulfonate.
 4. Acompound according to claim 3 wherein Y is bromo or iodo.
 5. A compoundaccording to claim 3 wherein Y is p-fluorobenzenesulfonate,trifluoromethanesulfonate or fluorosulfonate.
 6. A compound according toclaim 5 wherein Y is trifluoromethanesulfonate.
 7. A compound accordingto claim 1 wherein Y is a cyano group.
 8. A compound according to claim1 wherein Y is a carboxylic acid.
 9. A compound according to claim 1wherein Y is an N-aryl carboxylic amide derivative of Formula (A)##STR39##
 10. A compound according to claim 9 wherein:R₁ and R₃ are bothhydrogen; and R₂ is methoxy.